阿托伐他汀经ERK信号转导通路抑制CD40L诱导的内皮细胞E-选择素的表达

Inhibition of CD40L induced E-selectin expression in endothelial cells through extracellular signalregulated kinase signaling pathway

目的探讨阿托伐他汀对CD40L诱导人脐静脉内皮细胞（HUVEC）表达E-选择素的影响及其信号转导通路的关系。方法原代培养HUVEC，给予CD40L刺激和不同浓度阿托伐他汀干预。应用反转录聚合酶链反应（RT—PCR）和流式细胞术分别检测阿托伐他汀对CD40L诱导的HUVEC的E-选择素mRNA和细胞表面E-选择素表达情况。同时通过蛋白印迹法检测细胞外信号调节激酶（ERK1／2）蛋白磷酸化的表达。结果0．1、1、10μmol／L阿托伐他汀可浓度依赖性下调CD40L诱导的HUVECE-选择素mRNA及蛋白的表达。1μmol／L阿托伐他汀可使E-选择素蛋白下调48．68％，10μmol／L阿托伐他汀可使E-选择素蛋白下调70．25％。同时不同浓度阿托伐他汀均能抑制CD40L诱导的ERK1／2磷酸化水平，其表达分别下调至CIMOL刺激组的81％±6％、73％±5％和41％±5％。结论阿托伐他汀能通过ERK1／2信号转导通路抑制CD40L诱导的内皮细胞E-选择素表达。

Objective To investigate the regulatory effects of atorvastatin on CD40L induced E- seleetin expression in endothelial cells and the association thereof with signal pathway. Methods Human umbilical vein endothelial cells （HUVECs） were obtained from umbilical cord newly delivered and incubated with CD40L for 24 hours with or without pretreated by atorvastatin of the concentrations of 0. 1, 1, or 10 μmol/L. The protein and mRNA levels of E-selectin were detected by flow cytometry and reverse transcription polymerase chain reaction （RT-PCR） respectively. The extracellular signal regulated kinase （ERK） 1/2 activation was analyzed by Western blotting. Results The E-selectin mRNA expression level of the HUVECs treated by atorvastatin was lower than that of the CD40L stimulation group by 33.33% when the atorvastatin concentration was 1 μmol/L, and was lower by 45.16% when the atorvastatin concentration was 10 μmol/L. The E-selectin protein expression level of the HUVECs treated by atorvastatin was lower than that of the CD40L stimulation group by 48.68% when the atorvastatin concentration was 1 μmot/L, and was lower by 70. 25% when the atorvastatin concentration was 10 μmoL/L. The phosphorylation level of ERK1/2 was enhanced by CD40L stimulation and the CD40L induced phosphorylation of ERK1/2 decreased by 81% ±6% ,73% ± 5%, and 41% ±5% respectively after atorvastatin stimulation. Conclusion Atorvastatin decreases the CD40L induced E-selectin expression in endothelial cells while atorvastatin at 0. 1 - 10 μmol/L concentration-dependently through inhibiting the activation of ERK1/2.